Curcumin and o-Vanillin Exhibit Evidence of Senolytic Activity in Human IVD Cells In Vitro
Curcumin and o-Vanillin Exhibit Evidence of Senolytic Activity in Human IVD Cells In Vitro
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DOI：
10.3390/jcm8040433
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发表时间：
2019-04-01
影响因子：
3.9
通讯作者：
Haglund, Lisbet
中科院分区：
文献类型：
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作者：
Cherif, Hosni;Bisson, Daniel G.;Haglund, Lisbet
姜黄素和o-香兰素清除了衰老的椎间盘（IVD）细胞，减少了与炎症和背痛相关的衰老相关分泌表型（SASP）。来自退行性和非轻度退行性人类IVD的细胞来自器官捐献者和接受腰痛手术的患者。采用RT-qPCR检测离体细胞中衰老和SASP标志物的基因表达，免疫细胞化学（ICC）检测衰老、增殖和凋亡标志物的蛋白表达。采用酶联免疫吸附试验（ELISA）检测SASP因子的表达水平。通过红素- o染色和二甲基亚甲基蓝测定蛋白聚糖含量来验证基质的合成。采用Western blotting和ICC检测药物靶向的分子途径。我们发现，与来自非亲属个体的非轻度退变椎间盘相比，退变椎间盘的衰老细胞水平高出40%，而与来自同一个体的非轻度退变椎间盘相比，退变椎间盘的衰老细胞水平高出10%。较高的衰老水平与SASP升高有关。两种药物都清除了衰老细胞，治疗增加了退化ivd培养物中增殖细胞和凋亡细胞的数量。处理后SASP因子表达降低，基质合成增加。这些作用是通过Nrf2和NFkB途径介导的。
Curcumin and o-Vanillin cleared senescent intervertebral disc (IVD) cells and reduced the senescence-associated secretory phenotype (SASP) associated with inflammation and back pain. Cells from degenerate and non-mildly-degenerate human IVD were obtained from organ donors and from patients undergoing surgery for low back pain. Gene expression of senescence and SASP markers was evaluated by RT-qPCR in isolated cells, and protein expression of senescence, proliferation, and apoptotic markers was evaluated by immunocytochemistry (ICC). The expression levels of SASP factors were evaluated by enzyme-linked immunosorbent assay (ELISA). Matrix synthesis was verified with safranin-O staining and the Dimethyl-Methylene Blue Assay for proteoglycan content. Western blotting and ICC were used to determine the molecular pathways targeted by the drugs. We found a 40% higher level of senescent cells in degenerate compared to non-mildly-degenerate discs from unrelated individuals and a 10% higher level in degenerate compared to non-mildly-degenerate discs from the same individual. Higher levels of senescence were associated with increased SASP. Both drugs cleared senescent cells, and treatment increased the number of proliferating as well as apoptotic cells in cultures from degenerate IVDs. The expression of SASP factors was decreased, and matrix synthesis increased following treatment. These effects were mediated through the Nrf2 and NFkB pathways.